Tebuconazole-containing pesticide composition for controlling fusarium head blightand application thereof

ABSTRACT

A pesticide composition for controlling  Fusarium  head blight, which contains 1 to 150 parts by weight of jinggangmycin and 1 to 30 parts by weight of tebuconazole. The pesticide composition of the present invention has an obvious synergistic effect when used for controlling  Fusarium  head blight, can remarkably enhance the effect of controlling  Fusarium  head blight, and increase the ability of the human being in controlling  Fusarium  head blight of grain crops.

FIELD OF THE INVENTION

The invention belongs to the technical field of pesticide application.More specifically, the present invention relates to a pesticidecomposition for controlling Fusarium head blight, and further relates toan agricultural fungicide for controlling Fusarium head blight.

BACKGROUND OF THE INVENTION

Wheat and barley are the most important cultivated grain crops in theworld as well as one of the main food crops in China. Grain crops areoften confronted with the threat of fungal diseases such as smut, headblight, powdery mildew, rust, leaf blight, sheath blight and eye pot.These diseases may occur in worldwide grain growing regions, severelyaffecting the yield and quality of agricultural grain products.Especially, Fusarium head blight caused by Fusaria not only can resultin devastating yield loss, but also can produce toxins, such asdeoxynivalenol (DON) and zearalenon in infected grains, consequently,the quality of food, such as fermented food and brewed liquors, isseverely affected, and the health of the human being and animals areharmed.

The deoxynivalenol toxin includes DON, 3A DON and 15A DON toxins, whichhave acute and chronic toxicity to human being and animals. As the DONtoxin has stable chemical properties and won't be decomposed whenheated, the DON toxin widely exists in head blight fungus-infectedbarley and wheat, processed food thereof and animal products fed withgrains infected by head blight fungi. It has been known that the mainsymptoms of human and animal poisoning caused by the DON toxin includenausea, emesis, dizziness, drowsiness, headache, numb hands and feet,general weakness and decrease in immunity, the fluctuation of breath,pulse, body temperature and blood pressure, bleeding, miscarriage andeven death can be seen in severe cases, and the DON toxin can causehuman Kaschin-Beck disease (KBD) and has obvious embryotoxicity andteratogenic and carcinogenic effects. Since the DON toxin has severeharm to human health. In the joint conference on food additives andcontaminants held in Geneva in October, 1973, the United Nations worldfood and agriculture organization (FAO) and the world healthorganization (WHO) decided to include Fusarium toxins into the fifthtopic among sixteen topics given priority in research, and manycountries have also always considered the control of head blight as animportant research topic.

Moreover, many international organizations and national healthorganizations have laid down corresponding laws and regulations tostrictly limit DON toxin content in food. FAO provides that the contentof DON in food must be less than 1 mg/kg, and WHO provides that thecontent of the DON toxin in food and feed cannot respectively exceed thestandards of 1 mg/kg and 5 mg/kg. The European Union food safetystandard requires that the content of the DON toxin cannot exceed 0.75mg/kg in edible kernels and cannot exceed 0.5 mg/kg in bread and otherfoods. The national standard established by the Ministry of Health inChina provides that the content of the DON toxin in edible wheat flourand corn cannot exceed 1 mg/kg.

Fusarium head blight is an ascomycete disease, including bud rot,seedling blight and ear rot/ear blight, caused by a variety of Fusaria(Fusarium spp.). It includes nearly twenty varieties of pathogens, suchas Fusarium graminearum (F. graminearum), Fusarium asiaticum (F.asiaticum), Fusarium culmorum (F. culmorum), Fusarium avenaceum (F.avenaceum), Fusarium moniliforme (F. moniliforme) and Fusarium nivale(F. nivale). Fusarium head blight in most of the regions in China iscaused by the mixed population of Fusarium graminearum (F. graminearum)and Fusarium asiaticum (F. asiaticum), F. asiaticum is mainly in thesouthern wheat region, and F. graminearum is mainly in the northernwheat region.

As no grain crop varieties or gene resources which are immune or highlyresistant to Fusaria have been found as yet, using fungicides has becomethe only effective Fusarium head blight control technology. For almost acentury, people have been carrying out a great deal of work to researchand develop fungicides for grain fungal diseases, and have developedinorganic fungicides, organosulfureous fungicides and aromatichydrocarbon protective fungicides in succession, especially selectivefungicides developed after 1960s, such as benzimidazole fungicides,ergosterol biosynthesis inhibitors, cytochrome b inhibitors andsuccinodehydrogenase inhibitors, so that the epidemic harm of a lot ofimportant grain crop diseases, such as grain crop smut, powdery mildew,rust, leaf blight and eyespot, has been effectively controlled. However,as the deoxynivalenol (DON) toxin produced by Fusarium (Fusarium spp.)has the function of a pathogenic factor, the resistance to a variety offungicides is enhanced, and as a result, the control effect of themajority of fungicides on Fusarium head blight is not very ideal.Therefore, how to effectively control grain crop head blight is acurrent significant social demand in guaranteeing grain safety and foodsafety.

The DON toxin contamination level of diseased grain depends on thequantity of pathogenic fungi infecting the grain and the toxinbiosynthesis capability of thalli. Aimed at the situation that thecontrol effect of today's domestic and foreign main fungicides forcontrolling Fusarium head blight is not ideal and that DON toxincontamination often exceeds edible safety standards, the presentinvention studies and discovers two types of β-tubulin (β₁- and β₂-)receptors for benzimidazole fungicides existing in Fusaria and a geneticnegative regulation mechanism for fungicide resistance and fungicidesensitivity for the first time, and discovers that the 240th amino acidphenylalanine composing the β₂-tubulin receptor is a main cause offungicide sensitivity decrease and that the point mutation of the167^(th) amino acid will cause carbendazim to lose the control effectand greatly enhance the DON toxin synthesis capability, mycelialcolonization speed and pathogenecity of thalli, decreasing the controleffect of fungicides; in addition, the study also discovers anergosterol biosynthesis inhibitor (EBI) receptor of Fusarium, i.e.cytochrome, P₄₅₀ monooxygenase (Cyt.p₄₅₀), or fungicide sensitivity orfungicide resistance to EBI can be decreased by the overexpression orpoint mutation of encoding genes. Therefore, although some fungicidesshow very high activity in the lab, high dosages are needed when thefungicides are applied in fields, for example, 40 g to 50 g (600-750 ga.i./hm²) of benzimidazole fungicide with above active ingredients or 15g to 20 g (225-300 g a.i./hm²) of EBI with above active ingredients needto be used per mu to obtain about 70 percent of Fusarium head blightcontrol effect.

As these selective fungicides with single acting mechanisms are used ina large amount for a long term, fungicide resistance will appear inpathogenic fungus populations in the nature, and the effect in use willdecrease year by year. After nearly thirty years of fungicide resistancemonitoring, the inventor discovers that Fusaria which have developedfungicide resistance to benzimidazole fungicides, such as carbendazim,have formed a dominant population in Eastern China, and the commonlyused benzimidazole fungicides, such as carbendazim andthiophanate-methyl, have almost lost the value in controlling Fusariumhead blight. Furthermore, because the capability of fungicide-resistantpathogenic fungi in producing DON toxin is not less than five times thecapability of sensitive strains, the harm of Fusarium head blight andthe risk of food safety in China are increasingly exacerbated. In orderto reduce yield loss, farmers often double the use of fungicides, suchas carbendazim, and as a result, fungicide resistance, pesticideresidue, environment pollution and food safety problem are furtherexacerbated.

Therefore, the inventor finally accomplishes the present inventionthrough a great deal of experimental research and analysis on the basisof summarizing the prior art.

SUMMARY OF THE INVENTION Technical Problems to be Solved

The objective of the present invention is to provide a fungicidecomposition for controlling Fusarium head blight and reducing the dosageof conventional pesticides.

The other objective of the present invention is to provide anagricultural fungicide for controlling Fusarium head blight anddecreasing grain DON toxin contamination.

Technical Solution

The present invention is implemented through the following technicalsolution.

The present invention relates to a pesticide composition for controllingFusarium head blight.

The fungicide composition comprises jinggangmycin and tebuconazoleaccording to a weight ratio of (1-150):(1-30).

According to one preferred embodiment of the present invention, thepesticide composition comprises jinggangmycin and tebuconazole accordingto a weight ratio of (5-85):(3-24).

According to another preferred embodiment of the present invention, thepesticide composition comprises jinggangmycin and tebuconazole accordingto a weight ratio of (5-30):(6-18).

The present invention further relates to an agricultural fungicide forcontrolling Fusarium head blight. The agricultural fungicide contains0.1 to 90.0 percent by weight of fungicide composition, and the balanceis a carrier and/or an adjuvant acceptable in pesticides.

According to one preferred embodiment of the present invention, theagricultural fungicide contains 10 to 75.0 percent by weight offungicide composition, and the balance is the carrier and/or theadjuvant acceptable in pesticides.

According to another preferred embodiment of the present invention, theagricultural fungicide contains 26 to 58.0 percent by weight offungicide composition, and the balance is the carrier and/or theadjuvant acceptable in pesticides.

According to another preferred embodiment of the present invention, theagricultural fungicide is a suspension concentrate, an emulsion inwater, a microemulsion, a wettable powder or a water-dispersiblegranule.

According to another preferred embodiment of the present invention, thecarrier is one or more of water, attapulgite, kaolin and light calciumcarbonate.

According to another preferred embodiment of the present invention, theadjuvant is chosen from ethanol, methanol, ethylene glycol, propyleneglycol, NNO-1, NNO-7, xanthan gum, polyethylene glycol, glycerol, nekal,sodium dodecyl sulfate, sodium dodecyl benzene sulfonate, ammoniumsulfate, alkylphenol ethoxylates, alkylphenol polyoxyethylene etherphosphate, polyoxyethylene fatty acid, benzoic acid, sodium ligninsulfonate, carboxymethylcellulose and polyvinyl alcohol.

According to another preferred embodiment of the present invention, theFusarium head blight is wheat scab and barley scab, including bud rot,seedling blight, ear rot or ear blight of grain crops caused by Fusariumpathogenic fungi (Fusarium spp.) of carbendazim-resistant Fusaria.

The present invention is described in more detail below.

The present invention relates to an agricultural fungicide compositionfor controlling Fusarium head blight.

The pesticide composition comprises jinggangmycin and tebuconazoleaccording to a weight ratio of (1-150):(1-30).

The chemical structural formula of tebuconazole used in the presentinvention is as follows:

Tebuconazole is a highly effective, broad-spectrum and highly systemictriazole ergosterol biosynthesis inhibitor, which has three majorfunctions, including protection, treatment and eradication. Tebuconazoleis used as grain crop seed treatment agent and foliar spray worldwide,the fungicidal spectrum is wide, the fungicidal activity is high, andpersistence is long. The fungicide destroys the cell membranepermeability and membrane structure of fungi mainly by inhibitingergosterol biosynthesis, strongly inhibiting the growth of the fungalhyphae. However, not only do tebuconazole acceptors Cyt.P₄₅₀ indifferent plant pathogenic fungue cells have structural diversity, butalso a variety of regulatory and antagonistic physiological mechanismsexist, so that different fungi show different sensitivities. At present,this fungicide is mainly used for controlling a variety of fungaldiseases on crops including wheat, paddy rice, peanut, vegetables,banana, apple, pear, corn and broomcorn, and has been registered andused widely on more than sixty types of crops in more than fiftycountries around the world.

The chemical structural formula of jinggangmycin or validamycin used bythe present invention is as follows:

Jinggangmycin is a secondary metabolite of actinomycetes, and containssix types of similar amino glucan glycoside derivatives A, B, C, D, E,F. A large number of domestic and foreign studies have shown that thecomponent A is the main active component of jinggangmycin. Therefore,jinggangmycin in the present invention is jinggangmycin A.

Jinggangmycin shows interference specific to the development of hyphaeat the tips of rhizoctonia fungi (Rhizoctonia spp.) among basidiomycetesin vitro. Therefore, since jinggangmycin was discovered in 1970s,jinggangmycin has always served as an agricultural antibiotic forspecifically controlling plant diseases caused by rhizoctonia, or hasbeen processed into mixtures along with pesticides for controlling otherplant pests to control rhizoctonia (Rhizoctonia spp.) diseases. Theserhizoctonia diseases mainly include rice sheath blight, wheat sheathblight and sheath blight or damping-off of other crops. In recent years,it has also been discovered that jinggangmycin can also be used forcontrolling false smut caused by hard-to-culture imperfect fungi(Ustilaginoidea virens).

As validamycin or jinggangmycin only shows specific antifungal activityon rhizoctonia in vitro and does not have antifungal activity on Fusariaof head blight, people have always studied the selective mechanism ofjinggangmycin with rhizoctonia as an object for decades, discoveringthat validamycin/jinggangmycin interferes with the inositol andtrehalose metabolism of rhizoctonia, as a result, the structure of thecell wall is destroyed, rhizoctonia is prevented from infecting plants,and thereby jinggangmycin has a good protective effect. For decades,this antibiotic has always been limited to use for controllingrhizoctonia diseases of crops, especially rice sheath blight.

As everyone knows, fungicide-resistant pathogenic fungi can be easilyscreened out from specific fungicides with single effects, so thespecific fungicides will lose application value shortly after beingapplied widely. The inventor has studied the risk of the fungicideresistance of rhizoctonia to selective jinggangmycin since 1980s,discovering that what is different from any other selective antibioticsand chemical fungicides with extremely high resistance risk is thatjinggangmycin-resistant rhizoctonia cannot be screened out fromjinggangmycin under both lab and field conditions. It can be inferredtherefrom that besides interfering with known inositol and trehalosebiosynthesis, jinggangmycin may also have other specialdisease-resistant effect mechanisms.

On the basis of studying the DON toxin biosynthesis pathway andregulatory mechanism of Fusarium, the inventor carried out the screeningof a large number of compounds for inhibiting DON toxin biosynthesis. Inthe screening process, it was surprisingly discovered that with certaintreating dosage, jinggangmycin can strongly inhibit biochemical reactionin the early pathway of DON toxin biosynthesis of wheat scab fungi.Therefore, providing a technique for controlling Fusarium head blightwith a jinggangmycin and tebuconazole composition to reduce DONcontamination has become an important objective of the presentinvention.

The inventor carries out the inhibition of the synthesis of thepathogenic factor DON of Fusaria by jinggangmycin/validamycin and thescreening of synergistic formulas of mixtures ofjinggangmycin/validamycin and other various fungicides for controllingFusarium head blight in fields for the first time in the world, andcreatively discovered that the jinggangmycin and tebuconazolecomposition therein has the advantages of outstanding synergism, DONtoxin contamination reduction and great tebuconazole dosage reduction incontrolling Fusarium head blight.

In the pesticide composition of the present invention, if the amount ofjinggangmycin is within a range from 1 part by weight to 150 parts byweight while the amount of tebuconazole is less than 1 part by weight,then the control effect on head blight will be decreased; if the amountof tebuconazole is greater than 30 parts by weight, then cost will beincreased and the growth of wheat will be inhibited. Therefore, it isreasonable that the amount of tebuconazole is 1 to 30 parts by weight.

Likewise, if the amount of tebuconazole is within a range from 1 part byweight to 30 parts by weight while the amount of jinggangmycin is lessthan 1 part by weight, then the synergistic effect of jinggangmycin incontrolling head blight and decreasing the toxin contamination level incooperation with tebuconazole will be lost; if the amount ofjinggangmycin is greater than 150 parts by weight, then cost will beincreased, and the synergistic effect of jinggangmycin in controllinghead blight and reducing toxin contamination in cooperation withtebuconazole will be decreased; and therefore, it is appropriate thatthe amount of jinggangmycin is 1 to 150 parts by weight.

Preferably, the pesticide composition comprises jinggangmycin andtebuconazole according to a weight ratio of (5-85):(3-24).

More preferably, the pesticide composition comprises jinggangmycin andtebuconazole according to a weight ratio of (5-30):(6-18).

Both jinggangmycin and tebuconazole which are used by the presentinvention are conventional pesticide products which are sold in themarket at present, such as 3%, 5%A, 20%, 40%A and 60%A jinggangmycinpowders and aqueous solutions of Zhejiang Qianjiang Biochemical Co.,Ltd. and 25% tebuconazole wettable powder produced by Jiangsu YanchengShuangning Agrochemical Co., Ltd.

The present invention further relates to an agricultural fungicidepreparation for controlling Fusarium head blight. The agriculturalfungicide contains 0.1 to 90.0 percent by weight of jinggangmycin andtebuconazole fungicide composition, and the balance is a carrier and/oran adjuvant acceptable in pesticides.

In the present invention, it should be understood that the pesticidecarrier or adjuvant is an auxiliary substance which is used to improvethe physical and chemical properties of the agent, enhance efficacy andfacilitate transportation and storage when the pesticide preparation isprocessed or used, such as a wetting agent, an emulsifier, a dispersingagent, a binder, a stabilizer or a synergist. According to the presentinvention, any chemical substance which has these properties and doesnot have any adverse influence on the agricultural fungicide of thepresent invention can be used in the present invention, and they are allwithin the protection scope of the present invention.

The carrier used in the present invention, for example, is one or moreof water, attapulgite, kaolin and light calcium carbonate. Besides, thepresent invention can also use other carriers which can meet therequirement of the present invention and are commonly used in the art.

For example, the adjuvant used by the present invention is chosen fromethanol, methanol, ethylene glycol, propylene glycol, NNO-1 (chemicalname: naphthalene sulfonate formaldehyde condensate sodium salt), NNO-7(chemical name: naphthalene sulfonate formaldehyde condensate calciumsalt), xanthan gum, polyethylene glycol, glycerol, nekal, sodium dodecylsulfate, sodium dodecyl benzene sulfonate, ammonium sulfate, alkylphenolethoxylates, alkylphenol polyoxyethylene ether phosphate,polyoxyethylene fatty acid, benzoic acid, sodium lignin sulfonate,carboxymethylcellulose and polyvinyl alcohol. The adjuvant used by thepresent invention is a product which is sold widely in the market atpresent, such as dispersing agents sold with the product names “NNO-1”and “NNO-7” by Xinyi Feihuang Chemical Co., Ltd.,butylnaphtalenesulfonic acid sodium salt emulsifying, dispersing andpermeating agent sold with the product name “nekal” by Hubei ProsperityGalaxy Chemical Co., Ltd., alkylphenol ethoxylates solded by ShanghaiHonesty Fine Chemical Co., Ltd., alkylphenol polyoxyethylene etherphosphate sold by Guangzhou XiLu Chemical Co., Ltd., polyoxyethylenefatty acid sold by Shandong Liangshan Hongtai Epoxidized Soybean OilCo., Ltd. and carboxymethylcellulose sold by Jiangxi Pingxiang MashanAdditives Factory. Besides, the present invention can also use otheradjuvants which can meet the requirement of the present invention andare commonly used in the art.

In the present invention, in the agricultural fungicide, if the amountof the pesticide composition is less than 0.1 percent, then productionand transportation costs will be increased, the disease preventioneffect will be decreased or the synergistic effect will be lost; if theamount of the pesticide composition is greater than 90.0 percent, thenthe physical properties of the preparation will be deteriorated thesynergistic effect will be lost or the disease prevention effect will bedecreased ; and therefore, it is reasonable that the amount of thepesticide composition is 0.1 to 90.0 percent.

Preferably, the agricultural fungicide contains 10 to 75.0 percent byweight of pesticide composition, and the balance is the carrier and/orthe adjuvant acceptable in pesticides.

More preferably, the agricultural fungicide contains 26 to 58.0 percentby weight of pesticide composition, and the balance is the carrierand/or the adjuvant acceptable in pesticides.

In practical application, according to methods known by those skilled inthe art, the fungicidal composition of the present invention and thecarrier and/or the adjuvant can be prepared and processed into agentforms adopted commonly in agriculture, such as a suspension concentrate,an emulsion in water, a microemulsion, a wettable powder or awater-dispersible granule.

While the suspension concentrate is prepared,the applicable carrier oradjuvant,for example, can be a dispersing agent chosen from alkylnaphthalene sulfonate, polycarboxylate and lignosulfonate, a wettingagent chosen from alkylphenol polyoxyethylene ether phosphate, phenethylphenol polyoxyethylene ether phosphate, alkyl sulfate, alkyl sulfonateand naphthalene sulfonate, a thickening agent chosen from xanthan gum,aluminium-magnesium silicate and bentonite, a preservative chosen frombenzoic acid and sodium benzoate, an organosilicon defoaming agnt and anantifreezing agent chosen from glycerol, urea, ethylene glycol andpropylene glycol.

While the emulsion in water is prepared, the applicable carrier oradjuvant,for example, can be an emulsifier chosen from pesticideemulsifier 700, pesticide emulsifier 2201, Span-60 and emulsifier T-60,a solvent chosen from xylene, methylbenzene and cyclohexanone, astabilizer chosen from triphenyl phosphite and epichlorohydrin, anantifreezing agent chosen from ethylene glycol, propylene glycol,glycerol and urea, a thickening agent chosen from aluminium-magnesiumsilicate, bentonite and xanthan gum or a preservative chosen frombenzoic acid and sodium benzoate.

While the microemulsion is prepared, the applicable carrier oradjuvant,for example, can be an emulsifier chosen from calcium dodecylbenzene sulfonate (pesticide emulsifier 500), pesticide emulsifier 700,pesticide emulsifier 2201, Span-60, Tween-80 and TX-10, a cosolventchosen from methanol, isopropanol, n-butyl alcohol and ethyl alcohol, asolvent chosen from dimethylbenzene, methylbenzene, cyclohexanone andN-methylpyrrolidone or a stabilizer chosen from triphenyl phosphite andepichlorohydrin.

While the wettable powder is prepared, the applicable carrier oradjuvant,for example, can be a dispersing agent chosen frompolycarboxylate, lignosulfonate and alkyl naphthalene sulfonate, awetting agent chosen from alkyl sulfonate, alkyl sulfate and naphthalenesulfonate or a filler chosen from light calcium carbonate, talcumpowder, diatomite, kaolin and attapulgite.

While the water-dispersible granule is prepared, the applicable carrieror adjuvant,for example, can be a dispersing agent chosen frompolycarboxylate, lignosulfonate and alkyl naphthalene sulfonate, awetting agent chosen from alkyl sulfate, polyoxyethylene alcohol, alkylsulfonate and naphthalene sulfonate, a disintegrating agent chosen fromcitric acid, ammonium sulfate, glucose, urea and sodium bicarbonate, abinder chosen from corn starch, microcrystalline celluloses anddiatomite or a filler chosen from light calcium carbonate, sepiolite,diatomite, kaolin, attapulgite and talcum powder.

His research results are respectively elaborated below.

I. Screening of Active Compounds for Inhibiting Fusarium Growth

The present invention adopts a conventional method of fungicidebioassay, raw jinggangmycin and raw tebuconazole are respectivelyprepared into 2 mg/mL of mother solution by using sterile water andmethanol, a control agent, raw carbendazim, is dissolved into 0.1 M/L ofhydrochloric acid solution, phenamacril is dissolved in methanol, andthereby 2 mg/mL of mother solution is prepared. When potato dextroseagar media (PDA) are cooled to the temperature of about 45 □,jinggangmycin and tebuconazole are respectively added in to designedconcentrations (see Table 1), and are then poured into culture dishes toproduce plates treated by the different agents, the process is repeatedeach time after three dishes are treated, hyphal clumps of wildsensitive strains (sensitve strains for short) and carbendazim-resistantand phenamacril-resistant strains (fungicide-resistant strains forshort) of common Fusarium graminearum and Fusarium asiaticum causingFusarium head blight are inoculated, and are cultured under thetemperature of 25 □ for four days, the diameters of the fungal coloniesare measured by a crossing method, effective medium dosages (EC₅₀values) when different treatments inhibit the growth of the pathogenicfungi by 50 percent are calculated according to dosage-reaction curves,and antifungal activities are compared.

The experimental result indicates that no matter whether jinggangmycinis used alone or mixed with tebuconazole, jinggangmycin almost does nothave inhibitory activity on the growth of the sensitive strains andfungicide-resistant strains of the two types of Fusaria causing headblight in vitro, and only when the concentration of jinggangmycin is ashigh as 50 μg/mL can jinggangmycin have 6.5 to 7.8 percent of inhibitoryeffect on the growth of the two types of Fusaria. However, tebuconazolehas a similar strong inhibitory effect on the hyphal growth of thesensitive strains and fungicide-resistant strains of the two types ofFusaria, and the inhibitory effect of 0.078 μg/mL of tebuconazole fortreatment on hyphal growth can approximate 50 percent. Jinggangmycindoes not have a synergistic effect for tebuconazole in inhibiting hyphalgrowth in vitro (Table 1).

According to growth inhibition rates of the wild sensitive strains andcarbendazim-resistant and phenamacril-resistant strains of Fusariumasiaticum and Fusarium graminearum treated by different doses oftebuconazole, an effective medium dosage (EC₅₀) of tebuconazole forinhibiting the growth of the different fungicide-sensitive strains iscalculated, a result shows that the sensitivities of the sensitivestrains, the carbendazim-resistant strains and the phenamacril-resistantstrains to tebuconazole are similar, EC₅₀ is 0.085 to 0.116 μg/mL, and aexperimental result is listed in Table 2.

With EC₅₀ of the agents as parameters, the activities of the differentfungicides in inhibiting the growth of the Fusaria are compared, and itis discovered that the activity of tebuconazole is about 4.5 times theactivity (EC₅₀ is 0.45 μg/mL for both types of Fusaria) of carbendazimon the sensitive strains and about 1.5 times the activity (EC₅₀ is 0.165μg/mL for both types of Fusaria) of phenamacril on the sensitivestrains. The result indicates that tebuconazole has strong activity ininhibiting the growth of the wild sensitive strains and thecarbendazim-resistant and phenamacril-resistant Fusaria, helping todecrease the contamination level of DON toxin infecting grains andcontrol fungicide-resistant diseases.

TABLE 1 Influence of Jinggangmycin and Tebuconazole on Growth ofSensitive Strains of Two types of Fusaria In Vitro Average Fungal GrowthInhibition Strains Colony Diameter Rate Concentration (mm) (%) Agents(μg/mL) F.g* F.a* F.g F.a Jinggangmycin 0.1 76.3 76.3 / / A 1.0 76.276.3 / / 10.0 75.4 75.6 / / 50.0 70.3 69.4  6.6  7.8 Tebuconazole 0.0242 43 34.2 32.9 0.078 32 34 47.5 48.8 0.3125 21 22 72.1 70.8 1.25 16 1878.7 76.1 5.0 9 9 88.0 88.0 Jinggangmycin 0.1 + 0.078 30 31.5 60.1 58.1A + 1.0 + 0.078 32.8 31.8 56.4 57.7 Tebuconazole 10.0 + 0.078  29.8 32.460.4 56.9 50.0 + 0.078  30.5 30.2 59.5 59.9  0.1 + 0.3125 17.3 18.5 77.075.4  1.0 + 0.3125 18.2 18.1 75.8 75.9 10.0 + 0.3125 17.7 18 76.5 76.150.0 + 0.3125 16.1 15.9 78.6 78.9 Comparison 0 75.2 75.3 / / *F.g andF.a are abbreviations of Fusarium graminearum and Fusarium asiaticum,similarly hereinafter.

TABLE 2 Median Effective Concentration (EC₅₀) of Tebuconazole for GrowthInhibition of Sensitive Strains and Carbendazim-resistant andPhenamacril-resistant Strains of Two Types of Fusaria F.g F.g F.a F.aF.g carbendazim- phenamacril- F.a carbendazim- phenamacril- sensitiveresistant resistant sensitive resistant resistant Strains strainsstrains strains strains strains strains EC₅₀ (μg/mL) 0.096 0.101 0.0850.102 0.116 0.111 *F.g and F.a are abbreviations of Fusarium graminearumand Fusarium asiaticum, similarly herinafter.

II. Inhibitory Activity of Jinggangmycin on Toxin BiosynthesisCapability of Fusaria

As the fungicide sensitivities of Fusarium graminearum and Fusariumasiaticum to jinggangmycin and tebuconazole are the same, the inventorchooses carbendazim-resistant Fusarium asiaticum with higher toxinsynthesis capability (the weight of synthesized DON per unit fungusamount, μgDON/g by dry weight of hyphae) as a material to further studytoxin synthesis. Carbendazim-resistant Fusarium asiaticum causingFusarium head blight is inoculated into 3 percent of sterile mung beansoup, the solution is shaken for culture under the temperature of 25° C.and 12/24 hours of scattered light for ten days, and conidia arecollected centrifugally. The conidia are inoculated into potato sucrose(PS) culture solutions containing different doses of jinggangmycinaccording to final 10⁶/mL and shaken to be cultured under thetemperature of 25° C. and 12/24 hours of scattered light, cultures arefiltered after seven and fourteen days, toxin contents in the culturesolutions are assayed respectively and the dry weights of hyphae aremeasured, and toxin synthesis capabilities (the amount of toxin producedper unit weight of hyphae) are analyzed.

Toxin Determination Method: The culture filtrate is equal-volumeextracted with ethyl acetate for two times, the extracts are combined,and then depressurized, distilled and dried, the combined extract isdissolved by 1 mL of acetonitrile and transferred into a new centrifugetube, and after being distilled and dried again, the solution is storedunder −20 □ for later assay. During assay, 100 μL of TMS derivatizationreagent (TMSI:TMCS=100:1) is added, 1 mL of ultrapure water is addedafter 10 minutes of uniform mixing, supernate is extracted and addedinto a GC sample bottle after shaking for layering, and a gaschromatograph (GC-ECD) with an electron capture detector is used tocarry out toxin content assay. With a DON reagent of Sigma as a standardsample, a standard curve is created, and DON contents, including DON,3ADON and 15ADON, in the culture solutions are calculated. Meanwhile,the filtered hyphae are dried to constant weights under 80 □, and thedry weights of the hyphae are measured. In addition, after seven days ofshaking for culture, the hyphae are taken out, and the expression levelsof the key gene Tri5 for toxin synthesis are assayed.

It is discovered from an experimental result (Table 3) that the hyphalgrowths of the head blight fungi increase as culture time extends, but,when shaken to be cultured in the media treated by the different dosesof jinggangmycin, hyphal growths are not notably changed in comparisonwith that of the blank control. It is indicated that jinggangmycin doesnot have the inhibitory effect on the growth of the head blight fungicultured in liquid, which is the same as a linear growth ratedetermination result on the PDA plates. However, it is discovered forthe first time that the amount (μg DON/g by dry weight of hyphae) of DONtoxin synthesized per unit hyphal weight is remarkably decreased as thetreating dosage of jinggangmycin is increased, meeting the dosage effectlaw. Moreover, the inhibitory effect of jinggangmycin on DON synthesisdecreases as culture time extends, and especially, the decreasingamplitude of low-concentration treatment is greater. It is indicatedthat jinggangmycin may be degraded as experimental time extends, and asa result, the inhibitory effect on toxin biosynthesis may be decreased.

According to toxin synthesis gene expression level analysis after sevendays of treatment, it is creatively discovered that althoughjinggangmycin does not have adverse influence on the growth and hyphalmorphology of Fusaria in vitro, a very low treating dose ofjinggangmycin can strongly inhibit the expression of the key gene tri5for DON toxin synthesis, weakening the toxin biosynthesis capability ofthalli and reducing DON biosynthesis, and an experimental result islisted in Table 4.

TABLE 3 Effect of Jinggangmycin in Inhibiting DON Toxin SynthesisCapability of Fusaria Toxin Production Capability of Hyphae DONBiosynthesis Treating Dosage (μgDON/g by dry weight Inhibition Rate ofJinggangmycin of hyphae) (%) (μg/ml) 7 days 14 days 7 days 14 days 088.0 52.32 / / 1 62.82 51.43 28.61 1.70 10 56.76 48.20 35.50 7.87 10048.23 36.68 45.19 29.89 1000 30.35 22.52 65.51 56.96 10000 21.23 12.8575.88 75.44

TABLE 4 Influence of Jinggangmycin on Gene Expression of DON SynthesisGene Tri5 of Fusaria Rate of Inhibition on Treating Dosage RelativeExpression of Jinggangmycin Relative Expression Level of Tri5 Gene(μg/ml) Level of Tri5 Gene (%) 0 1 / 1 0.41 59 10 0.34 66 100 0.16 841000 0.13 87 10000 0.10 90

III. Synergistic Effect of Tebuconazole for Jinggangmycin in InhibitingDON Toxin Biosynthesis Capability of Fusaria

Tebuconazole treatment can destroy the cell membrane permeability of thehead blight fungi, inhibiting the growth of hyphae. Tebuconazole andjinggangmycin composition treatment can enhance the absorption andutilization of jinggangmycin by the pathogenic fungi. While the researchcontent II determines the inhibition of jinggangmycin on DON toxinbiosynthesis of the Fusaria, inhibitory effects of jinggangmycin on theDON toxin biosynthesis capability of Fusarium asiaticum under theexistence of 0.1 μg/mL of tebuconazole are determined after seven andfourteen days of culture, and the synergistic effect of tebuconazole forjinggangmycin in inhibiting DON toxin biosynthesis is analyzed. The DONassay method is the same as that of the research content II.

It has been known from the research contents I and II that 0.1 μg/ml oftebuconazole for treatment alone has an inhibition rate of about 50percent on the hyphal growth of the head blight fungi. It can be seenfrom the result in Table 5 that 0.1 μg/ml of tebuconazole does not havesignificant inhibitory effects on the toxin biosynthesis capability ofthe Fusaria in comparison with the toxin biosynthesis capability of theblank control after seven and fourteen days of treatment, whichindicates that tebuconazole only has hyphal growth inhibition activity,and has no inhibitory effect on toxin synthesis capability.Nevertheless, when 0.1 μg/ml of tebuconazole exists in each treatmentconcentration, the inhibitory effect of jinggangmycin on DON toxinbiosynthesis capability is greatly enhanced. Moreover, as culture timeextends, the decreasing speed of the inhibitory effect of jinggangmycinon toxin synthesis capability is remarkably lower than that of controltreatment without tebuconazole, and in particular, the synergisticeffect for low-concentration jinggangmycin treatment and the prolongingof action time are more obvious. Based on the results of thesimultaneous experiments in Table 3, synergistic effects forjinggangmycin in inhibiting the toxin biosynthesis of the Fusaria afterseven and fourteen days of treatment under the existence of 0.10 μg/mLof tebuconazole are calculated, and results are listed in Table 5. Theseresults indicate: (1) jinggangmycin has a strong effect in decreasingthe DON toxin biosynthesis capability of the Fusaria, while tebuconazoledoes not have this effect; (2) tebuconazole has an obvious synergisticeffect for jinggangmycin in inhibiting the toxin biosynthesis of theFusaria, and as the treating dosage of jinggangmycin is reduced, thesynergistic effect is enhanced; (3) tebuconazole can prolong the time ofthe inhibitory effect of jinggangmycin on the toxin synthesis of theFusaria, and the synergistic effect is enhanced as the time of treatmentextends.

TABLE 5 Synergistic Effect of 0.1 μg/mL of Tebuconazole forJinggangmycin in Inhibiting DON Toxin Synthesis of Wheat Srab FungiSynergistic coefficient of DON 0.1 μg/ml of Toxin ProductionBiosynthesis Tebuconazole for Capability Inhibition Jinggangmycin inTreating Dosage (μgDON/g by dry Rate Inhibiting DON of Jinggangmycin +weight of hyphae) (%) Synthesis * Tebuconazole 7 14 7 14 7 14 (μg/ml)days days days days days days 0 + 0  88.00 52.32 / / / / 0 + 0.1 87.4252.50 4.2 0.6 / / 1 + 0.1 47.20 45.15 45.1 17.6 157.6 1034.4 10 + 0.1 26.06 27.21 68.6 50.5 193.1 641.9 100 + 0.1  15.13 10.08 83.7 78.5 185.3262.7 1000 + 0.1   8.65 5.82 89.5 87.8 136.6 154.2 10000 + 0.1   5.853.12 92.8 92.4 122.3 122.5 * Synergistic Coefficient Calculation Method:When 0.1 μg/ml of tebuconazole exists, an inhibition rate ofjinggangmycin on toxin synthesis is divided by a toxin synthesisinhibition rate of a corresponding dose of jinggangmycin for treatmentalone, and the result is multiplied by 100.

IV. Effect and Synergistic Effect of Jinggangmycin and TebuconazoleComposition Suspension Concentrate in Treating Seeds to Control HeadBlight and Other Diseases at Wheat Seedling Stage

According to weight ratios of jinggangmycin to tebuconazole which are1:1.5, 10:1.5, 100:1.5, 1:1, 10:1, 100:1, 2:1, 20:1, 200:1, 5:1, 50:1and 500:1, composition suspension concentrate samples, the contents ofwhich are 3% (1:1.5, 1:1, 2:1), 5% (10:1.5, 10:1, 5:1), 10% (50:1,100:1, 20:1) and 20% (100:1.5, 200:1, 500:1), are prepared respectively.In a sample preparation method, raw tebuconazole and raw jinggangmycinare mixed according to the above-mentioned weight ratios, added into asolution containing 40 percent by weight of water, 8 percent by weightof ethylene glycol and propylene glycol mixed solvent (volume ratio:1:1), 1.0 percent by weight of NNO-1 and 1.5 percent by weight of NNO-7dispersing agents (produced by Xinyi Feihuang Chemical Co., Ltd.), 0.5percent by weight of xanthan gum binder and 1 percent by weight ofpolyethylene glycol and, finally, supplemented with water to 100percent, and the granule diameter of agents milled to 90 percent by asand mill is less than or equal to 5 μm.

According to dosages of the single agents in a mixed agent for treatmentper 100 kg of wheat seeds, single dosages of jinggangmycin andtebuconazole in corresponding dosages are designed (see Table 6), andbefore seeding, the samples are respectively mixed with the seedsaccording to 5 L of liquor per 100 kg of seeds to treat the wheat seedsinfected by wheat scab fungi after being diluted by water. Twenty fiveseeds are sowed per pot with the diameter of 20 cm, each treatment isrepeated for ten pots, and the seeds are put into a greenhouse forculture. Ten days after emergence, the rate of emergence and the rate ofdead seedlings are checked, and control effect on bud rot and seedlingblight caused by the head blight fungi is calculated. Meanwhile, tenseedlings which are uniform in growth are kept, furthermore, a branmedium containing rhizoctonia cerealis is inoculated in substrate,powdery mildew fungus and leaf rust fungus spores are inoculated onleaves, one hundred wheat seedlings per treatment are investigatedfourteen days after inoculation, and incidences of sheath blight,powdery mildew and rust and control effect are calculated. Meanwhile,according to prevention effects of the single agents on bud rot andseedling blight caused by the head blight fungi, theoretical preventioneffect (E=X+(100−X)Y/100, wherein E is the theoretical preventioneffect, X is the prevention effect of the single jinggangmycin agent,and Y is the prevention effect of the single tebuconazole agent) andsynergistic coefficient (actual prevention effect of the composition inapplication÷theoretical prevention effect'100) of the composition arecalculated by the Abbott (1925) method, and results can be seen in Table6.

TABLE 6 Effect and Synergistic Effect of Jinggangmycin and TebuconazoleComposition Suspension Concentrate in Treating Seeds to Control WheatDiseases Treating Dosage Control of Head Blight Per 100 kg of SeedsIncidence Prevention Effect % Theoretical Synergistic Serial (g ai.agent) Head Sheath Powdery Prevention Coefficient Number JinggangmycinTebuconazole Blight Blight Mildew Rust Effect % *  1 10 15 99.56 97.6591.25 96.32 70.61 141.00  2 100 15 99.21 99.35 93.22 97.15 74.72 132.77 3 1000 15 98.92 100 91.24 98.22 76.63 129.08  4 10 10 96 96.32 86.2296.2 60.85 157.76  5 100 10 95.23 98.22 82.14 95.32 66.33 143.57  6 100010 98.56 99.36 83.54 96.28 68.88 143.10  7 10 5 86.59 82.65 76.23 90.2651.35 168.63  8 100 5 90.64 89.25 76.25 87.96 58.16 155.84  9 1000 595.15 94.66 75.35 89.63 61.32 155.16 10 10 2 36.5 62.35 58.66 69.5 21.09173.07 11 100 2 56.84 72.66 59.25 83.64 32.14 176.87 12 1000 2 61.5480.95 62.54 85.36 37.27 165.13 13 10 0 0 10.5 0 0 / / 14 100 0 14.0 26.00 0 / / 15 1000 0 20.5 56.2 6.5 4.5 / / 16 0 15 70.61 42.55 86.5 86.35 // 17 0 10 60.85 21.61 78.3 81.24 / / 18 0 5 51.35 16.85 62.5 70.21 / /19 0 2 21.09 12.35 39.55 53.66 / / 20 Blank Control 16.5 16.5 21.5 65 24/ Incidence %

*Abbott believed that if the ratio of the actual prevention effect totheoretical prevention effect of a composition is greater than 1 (or asynergistic coefficient is greater than 100), then the composition has asynergistic effect (similarly hereinafter).

The above-mentioned experimental result indicates that when being mixedwith the wheat seeds to treat the wheat seeds infected by head blight,the jinggangmycin and tebuconazole composition not only can effectivelycontrol bud rot and seedling blight caused by the head blight fungi, butalso has an excellent synergistic effect in controlling seedling blightcaused by the head blight fungi within the ranges of the experimentaldosages for treatment (10-1000 g ai of jinggangmycin and 2-15 g ai oftebuconazole/100 kg of seeds), the synergistic coefficient is fargreater than 100, and the synergistic effect is extremely outstanding.The single jinggangmycin agent has a certain prevention effect on sheathblight, but is almost ineffective to powdery mildew and rust, while thesingle tebuconazole agent has a good prevention effect on head blight,sheath blight, powdery mildew and rust at the seedling stage, and canshow an obvious synergistic effect when mixed with jinggangmycin.

V. Synergistic Effect and Toxin-Reducing Effect of Jinggangmycin andTebuconazole Composition Suspension Concentrate in Controlling WheatScab

According to weight ratios of jinggangmycin to tebuconazole which are1:50, 1:20, 1:3, 1:2, 1:1.5, 1:1, 2:1, 5:1, 5:1.5, 50:3, 10:1, 10:1.5,50:2.5, 50:1.5 and 50:1, experimental composition suspension concentratesamples, the contents of which are 20% (1:50, 1:20, 1:3, 1:2, 1:1.5,1:1, 2:1, 5:1, 5:1.5) and 40% (10:1, 10:1.5, 50:3, 50:2.5, 50:1.5, 50:1)by weight, are prepared respectively. In a sample preparation method,after raw tebuconazole and raw jinggangmycin are mixed according to theabove-mentioned weight ratios, each mixture is added into a solutioncontaining 5 percent by weight of ethylene glycol and propylene glycolmixed solvent (volume ratio: 1:1), 2 percent by weight of glycerol, 1.0percent by weight of NNO-1, 1.5 percent by weight of NNO-7 dispersingagent, 3 percent by weight of sodium dodecyl sulfate and a small amountof water, finally, the solution is supplemented with water to 100percent, and the granule diameter of agents milled to 90 percent by asand mill is less than or equal to 5 μm.

Different dosages of the jinggangmycin and tebuconazole composition usedper mu are designed, and meanwhile, corresponding single-agenttreatments are designed according to dosages of the single agents ineach mixed agent in use. In April, 2013, experimental screening forcontrolling head blight in fields was carried out in Jiangsu White HorseLake Farm, the wheat variety was Huaimai 22, each agent sample was mixedwith water and sprayed for treatment at the initial stage of wheatblooming, five days later the agent sample is sprayed for the secondtime, and the amount of sprayed water per mu was 50 kg. Each treatmentwas repeated for three plots, and the area of each plot was 50 squaremeters. Meanwhile, 80 g of carbendazim wettable powder which was 50percent was used as a control agent for treatment per mu. According to acorresponding method specified by the industrial standard of Guidelinesfor the Field Efficacy Trials of Fungicides issued by the Ministry ofAgriculture, the occurrence of wheat scab was investigated at the stageof milky ripeness, and according to the actual effect of each treatmentin controlling head blight, the synergistic effects of the compositionwere calculated. By the Abbott (1925) method, theoretical preventioneffect (E=X+(100−X)Y/100, wherein E is the theoretical preventioneffect, X is the prevention effect of the single jinggangmycin agent,and Y is the prevention effect of the single tebuconazole agent) andsynergistic coefficient (actual prevention effect of the composition inapplication÷theoretical prevention effect×100) of the composition werecalculated.

Toxin Determination Method: Two hundred wheat ears were sampled everyfive treated points at the stage of wax ripeness, and were threshedindoors, and after drying, 30 g of kernels were sampled randomly andground. According to Goswami and Kistler methods, 5 g of flour was putinto a centrifuge tube, 20 mL of extract of acetonitrile and water(84:16) was added into the flour and uniformly mixed by a vortex mixer,the solution was then shaken on a shaker for 24 hours, and wascentrifuged at 5000 rpm for 10 minutes, and 2 mL of supernate wasblow-dried by nitrogen in an Eppendorf centrifuge tube and preservedunder −20 □. During assay, 100 82 L of TMS derivatization reagent(TMSI:TMCS=100:1) was added, 1 mL of ultrapure water was added after 10minutes of uniform mixing, supernate was extracted and added into a GCsample bottle after shaking for layering, and a gas chromatograph(GC-ECD) with an electron capture detector was used to carry out toxincontent assay.

Kernel-inflecting Fungus Amount Determination Method: 2 g of flour fortoxin assay was put into a 50 mL centrifuge tube, added with 20 percentof CTAB pathogenic fungus DNA extraction buffer, then added with 50 μLof protease K and 30 μL of RNA enzyme and mixed sufficiently, thesolution was centrifuged at 10000 rpm for 10 minutes after beingincubated under 65 □ for 3 hours, 20 mL of supernate was completelytransferred into a 50 mL centrifuge tube, then added with phenol,chloroform and isoamyl alcohol (25:24:1) of the same volume, shakenviolently and centrifuged at 10000 rpm for 5 minutes, 10 mL of supernatewas added with 3molL⁻¹ of cold sodium acetate accounting for one tenthof the volume and absolute ethyl alcohol two times the volume,precipitation was carried out under −20 □ for 24 hours, centrifugationwas carried out at 10000 rpm for 10 minutes, precipitate was eluted twotimes by 20 mL of 70 percent ethyl alcohol, the precipitate wasdissolved into 500 μL of TE solution after being dried, and the DNAtemplate was stored under −20 □. Primers were designed according to thekey gene Tri5 for DON synthesis, DNA in a sample is amplified byadopting real time quantitative PCR, moreover, the Tri5 DNA content(μgDNA/g of wheat) in each gram of wheat sample was calculated, and theinhibitory effect of each treatment of the composition on the toxinsynthesis capability of thalli was calculated. The result can be seen inTable 7.

TABLE 7 Synergistic Effect of Jinggangmycin and Tebuconazole CompositionSuspension Concentrate in Controlling Wheat Scab and Effect ofJinggangmycin and Tebuconazole Composition Suspension Concentrate inReducing DON Toxin Contamination DON Grain Disease Index Synthesis ToxinPrevention Effect % Capability Content DON Treating Dosage ActualTheoretical (μg of (μg of Contamination Serial (g ai. agent/mu)Prevention Prevention Synergistic toxin/g of toxin/g of Reduction NumberJinggangmycin Tebuconazole Effect Effect Coefficient DNA) kernels) Rate%  1 0.5 25 98.23 86.31 113.81 20.60 0.26 97.55  2 1 20 92.30 76.54120.59 19.65 0.34 96.79  3 5 5 77.35 47.78 161.89 14.85 0.82 92.26  4 510 89.58 59.15 151.44 14.98 0.28 97.36  5 5 15 96.20 74.31 129.46 15.060.21 98.02  6 10 3 58.23 43.14 134.99 8.86 1.51 85.75  7 10 5 79.6949.94 159.56 8.34 0.43 95.94  8 10 10 95.32 60.84 156.66 8.06 0.32 96.98 9 10 15 96.88 75.37 128.54 8.36 0.27 97.45 10 50 3 76.28 51.23 148.895.92 1.10 89.62 11 50 5 87.62 57.07 153.53 5.86 0.37 96.51 12 50 1095.18 66.42 143.30 5.68 0.20 98.11 13 50 15 97.25 78.88 123.29 5.87 0.1898.30 14 100 3 88.15 55.15 159.84 4.21 0.65 93.87 15 100 5 92.86 60.51153.45 4.12 0.21 98.02 16 100 10 94.84 69.11 137.22 3.86 0.23 97.83 17100 15 98.20 80.57 121.88 3.58 0.20 98.11 18 150 3 88.29 60.59 145.723.10 0.31 97.08 19 0.5 / 0.00 / / 22.51 9.40 11.32 20 1 / 0.00 / / 21.658.90 16.04 21 5 / 10.60 / / 16.85 8.60 18.87 22 10 / 14.30 / / 9.21 6.6037.74 23 50 / 26.50 / / 6.25 4.70 55.66 24 100 / 32.40 / / 4.25 4.5057.55 25 150 / 40.60 / / 3.01 4.00 62.26 26 / 3 33.65 / / 19.88 5.8045.28 27 / 5 41.59 / / 18.35 4.90 53.77 28 / 10 54.31 / / 18.95 4.2060.38 29 / 15 71.26 / / 19.02 3.60 66.04 30 / 20 76.54 / / 19.41 1.9082.08 31 / 25 86.31 / / 18.95 0.42 96.04 32 Control Agent: 67.5 / /28.94 4.40 58.49 Carbendazim 40 g a.i. 33 Blank Control 47.2 / / 21.5110.60 / Disease Index

The field screening and experiment results of the synergistic effect ofthe composition for controlling Fusarium head blight in the presentinvention indicate that when the jinggangmycin and tebuconazolecomposition is sprayed from the initial stage of wheat blooming to thefilling stage and the mixed agent of 1 g to 150 g of activejinggangmycin ingredient and 3 g to 25 g of active tebuconazoleingredient is used per mu, not only does the jinggangmycin andtebuconazole composition have an outstanding synergistic effect (thesynergistic coefficient is greater than 100) in controlling head blight,but also, after being applied, the jinggangmycin and tebuconazolecomposition has a remarkable synergistic effect in decreasing the DONcontamination level of grains, the DON content can be reduced by 85percent or above, and the DON contamination level can be controlled at asafe level not higher than 1 mg of DON/kg of grain. Meanwhile, theresult of quantitative pathogenic fungus analysis for the inflectedkernels also indicates that jinggangmycin has the effect of inhibitingthe toxin synthesis capability of thalli while tebuconazole does nothave such an effect. Although carbendazim has 67.5 percent of preventioneffect on head blight when used for treatment alone, the toxincontamination level of grains is still 4.4 mg/kg of grains, and is onlydecreased by 58.5 percent in comparison with the blank control.

VI. Synergistic Effect of Jinggangmycin and Tebuconazole CompositionSuspension Concentrate in Controlling Wheat Diseases

Adopting the experimental sample preparation methods of the inventionresearch experiments IV and V, the present invention further designstreating dosages of the composition in use, and also designssingle-agent treatments according to the dosages of jinggangmycin andtebuconazole in the composition. A field experiment was carried out inJiangsu Xinyang Farm in 2013. Each agent was mixed with water andsprayed to treat Huaimai 22 at the initial stage of wheat blooming, theagent was sprayed for the second time 5 days later, and the amount ofsprayed water per mu was 50 kg. Each treatment was repeated three times,and the area of each plot was 50 square meters. Meanwhile, 80 mL ofcarbendazim suspension concentrate which was 50 percent was used as acontrol agent for treatment per mu. According to a corresponding methodspecified by the relevant industrial standard of Guidelines for theField Efficacy Trials of Fungicides issued by the Ministry ofAgriculture, the occurrences of wheat scab, powdery mildew, rust andleaf blight were investigated at the stage of milky ripeness, diseaseindexes, control effect and synergistic effect for the control of headblight calculated by the above-mentioned Abbott (1925) method werecalculated, and the toxin assay method was the same as the method of theabove-mentioned research V (results can be seen in Table 8).

TABLE 8 Effect of Jinggangmycin and Tebuconazole Composition SuspensionConcentrate in Controlling Wheat Diseases and Reducing ToxinContamination of Grains Theoretical Synergistic Prevention CoefficientGrain Effect of Toxin of Mixed Content Mixed agent (μg Disease IndexPrevention Agent in of DON Thousand Treating Dosage Effect % onControlling toxin/g Reduction Kernel Serial (g ai. agent/mu) Head LeafPowdery Head Head of Percentage Weight Number Jinggangmycin TebuconazoleBlight Blight Mildew Rust Blight blight kernels) (%) (g)  1 1 15 92.6592.54 86.31 97.68 82.69 112.04 0.26 97.94 44.22  2 3 10 93.25 89.3581.22 91.52 68.25 136.63 0.36 97.14 44.16  3 5 5 80.23 82.15 76.51 85.6452.91 151.63 1.25 90.08 44.19  4 5 10 82.69 87.65 80.31 89.24 70.03118.08 0.84 93.33 44.42  5 5 15 93.15 88.25 84.54 96.41 83.66 111.340.62 95.08 44.35  6 10 3 76.52 76.54 76.54 76.54 48.26 158.57 1.85 85.3243.81  7 10 10 87.56 88.21 81.24 86.24 72.47 120.82 0.70 94.44 44.67  810 15 92.34 95.78 86.24 91.52 84.99 108.65 0.56 95.56 45.56  9 50 382.45 83.25 70.51 85.34 55.90 147.50 0.78 93.81 44.78 10 50 10 91.2487.15 81.32 89.21 76.54 119.21 0.35 97.22 44.67 11 50 15 95.31 97.2884.65 98.66 87.21 109.29 0.28 97.78 45.18 12 100 5 83.65 86.54 81.6683.08 66.18 126.40 0.42 96.67 45.22 13 100 10 92.84 91.26 76.55 88.1578.47 118.31 0.31 97.54 45.59 14 100 / 32.20 8.30 7.50 4.90 / / 3.3673.33 43.2 15 50 / 26.10 7.80 4.50 3.80 / / 3.8 69.84 42.8 16 10 / 13.307.80 0.00 0.00 / / 4.98 60.48 42.9 17 5 / 5.60 0.00 0.00 0.00 / / 7.9237.14 42.6 18 3 / 0.00 0.00 0.00 0.00 / / 8.98 28.73 42 19 1 / 0.00 0.000.00 0.00 / / 11.05 12.30 39.6 20 / 15 82.69 76.60 78.70 83.80 / / 1.6886.67 43.44 21 / 10 68.25 71.10 74.30 78.40 / / 2.51 80.08 42.78 22 / 550.12 52.10 70.20 66.70 / / 3.54 71.90 42.14 23 / 3 40.32 38.20 48.6041.90 / / 4.05 67.86 40.24 24 Carbendazim 38.60 48.50 52.60 61.20 / /5.67 55.00 41.60 40 g a.i. 25 Control 28.10 5.20 18.50 16.90 / / 12.60 /38.90 Disease Index

The field application results of the above-mentioned mixed agentsindicate that when 1 g to 100 g of jinggangmycin as an active ingredientand 3 g to 15 g of tebuconazole as an active ingredient are used per muto control wheat scab, the jinggangmycin and tebuconazole compositionsuspension concentrate not only has an excellent control effect on wheatscab, leaf blight, powdery mildew and rust, which is far better thanthat of commonly used carbendazim (the assay showed that pathogenicfungi which showed resistance to carbendazim in pathogenic funguscolonies in the experimental field accounted for 37.7 percent),moreover, the jinggangmycin and tebuconazole composition suspensionconcentrate has extremely remarkable synergistic and control effects onhead blight caused by fungicide-resistant head blight fungi, and theresult was similar to the field experiment result of White Horse LakeFarm in the same year. The difference between the effect of 3 g to 50 gof jinggangmycin and 3 g to 15 g of tebuconazole used per mu and theprevention effect of higher dosages of jinggangmycin is not great, DONtoxin contamination can be controlled within a safe range, moreover, thedosage is less, the cost is low, the environment can be protected, andbenefits are better. It is particularly worth noting that the thousandkernel weights of grains treated by all fungicides are obviously higherthan that of the control, especially the jinggangmycin and tebuconazolecomposition for treatment increases thousand kernel weight by not lessthan 14 percent, and the yield-increasing effect is more prominent.

VII. Synergistic Effect of Jinggangmycin and Tebuconazole CompositionWettable Powder in Controlling Wheat Scab and ToxinContamination-reducing Effect of Jinggangmycin and TebuconazoleComposition Wettable Powder

According to the good result of the jinggangmycin and tebuconazolecomposition suspension concentrate in the field control of head blightand the treatment of other diseases, the present invention designsweight proportions which can help to reduce the dosage of the pesticideand cost, and prepares composition wettable powders with differentcontents of jinggangmycin and tebuconazole. Preparation method: Aftermixing according to a weight ratio (percentage) of raw tebuconazole toraw jinggangmycin, the mixture is added with 30 percent by weight ofattapulgite carrier, 1.0 percent by weight of NNO-1 dispersing agent, 3percent by weight of nekal wetting agent and 2 percent by weight ofsodium dodecyl benzene sulfonate surfactant and, finally, added withlight calcium carbonate as a filler to 100 percent, and the mixture ismilled by a sand mill and screened by a 300-mesh screen. Compositionwettable powder samples, the contents of which are 20% (1:3, 1:2, 1:1.5,1:1) and 60% (5:1, 10:3, 10:1), are prepared respectively.

Meanwhile, according to the dosages of the single agents in thecomposition used per mu, the single-agent treatments of jinggangmycinand tebuconazole and a common polyketone wettable powder control agentare designed. In 2014, each composition wettable powder sample was mixedwith water and sprayed for treatment at the initial stage of wheatblooming, the composition wettable powder sample was sprayed for thesecond time 5 days later, and the amount of sprayed water per mu was 50kg. Each treatment was repeated three times, and the area of each plotwas 50 square meters. According to a corresponding method specified byGuidelines for the Field Efficacy Trials of Fungicides, the occurrencesof barley scab, powdery mildew, rust and leaf blight were investigatedat the stage of milky ripeness, and disease indexes and control effectwere calculated. Meanwhile, according to the Abbott (1925) method in theresearch content V, the synergistic effects of the mixed agents incontrolling barley scab were calculated. One hundred and fifty wheatears were sampled randomly every five treated points at the stage of waxripeness, taken and threshed indoors and assayed according to the methodof the research content V, and toxin contents of the kernels werecalculated. The treating dosages, prevention effects, synergisticeffects and influences of the agents on toxin content can be seen inTable 9.

TABLE 9 Effect of Jinggangmycin and Tebuconazole Composition WettablePowder in Controlling Barley Scab Treating Control of Head DosageDisease Index Prevention Blight Grain Toxin Toxin (g ai. Effect %Theoretical Content (mg Reduction Serial agent/mu) Head Leaf PowderyPrevention Synergistic of DON/Kg Percentage Number JinggangmycinTebuconazole Blight Blight Mildew Rust Effect Coefficient of grain) (%) 1 3 10 81.42 79.38 76.36 84.35 62.51 130.25 1.25 85.35  2 3 20 93.5284.63 90.48 92.08 78.40 119.29 0.38 95.55  3 5 15 85.21 82.54 85.2194.35 69.05 123.41 0.95 88.86  4 5 10 81.32 79.51 81.24 89.51 64.20126.67 0.85 90.04  5 5 5 82.16 69.28 86.24 80.65 49.88 164.71 0.84 90.15 6 10 15 89.33 85.21 81.64 90.84 71.67 124.63 0.64 92.50  7 10 10 86.2177.65 84.21 86.54 67.23 128.22 0.65 92.38  8 10 3 76.24 63.44 71.3675.34 39.03 195.34 0.78 90.86  9 50 10 89.51 76.14 83.22 87.29 72.41123.62 0.55 93.55 10 100 10 89.21 85.21 83.24 91.24 74.69 119.43 0.5493.67 11 3 / 0 0 0 0 / / 6.67 21.81 12 5 / 4.5 0 0 0 / / 5.28 38.10 1310 / 12.6 0 0 0 / / 4.62 45.84 14 50 / 26.4 0 0 0 / / 3.86 54.75 15 100/ 32.5 6.9 3.4 0 / / 3.49 59.09 16 / 3 30.24 34.61 48.69 68.2 / / 3.6557.21 17 / 5 47.52 51.24 63.54 76.54 / / 3.98 53.34 18 / 10 62.51 63.4172.61 77.24 / / 2.25 73.62 19 / 15 67.59 72.64 86.24 86.34 / / 1.7479.60 20 / 20 78.40 75.44 88.13 90.06 / / 1.23 85.58 21 Polyketone 67.362.60 74.12 73.31 / / 5.86 31.30 30 g a.i. 22 Control 20.5 25.30 14.438.62 / / 8.53 / Disease Index

The field application of the composition indicates that the single-agenttreatment of the single tebuconazole agent has a good prevention effecton barley scab, leaf blight, powdery mildew and rust while, excepthaving a low prevention effect on head blight, the single jinggangmycinagent almost has no effect on other diseases. Nevertheless, when thecomposition containing 3 g to 50 g ai of jinggangmycin and 3 g to 15 gai of tebuconazole is used, the prevention effect on head blight, leafblight, powdery mildew and rust is greatly enhanced, the control effectreaches not less than 80 to 90 percent, better than the effect of thecommon control agent polyketone, and the synergistic effect isoutstanding. In particular, when 3 g to 50 g of active jinggangmyciningredient and 3 g to 15 g of active tebuconazole ingredient are usedper mu, the dosage of the tebuconazole chemical fungicide can be greatlyreduced, the environmental pressure caused by pesticides can bedecreased, and the toxin contamination of kernels can be reduced.

VIII. Effect of Jinggangmycin and Tebuconazole Wettable Powder TankMixture in Controlling Wheat Diseases and Reducing Toxin Contamination

64 percent of jinggangmycin A soluble powder of Zhejiang Tonglu HuifengBiochemical Co., Ltd and 20 percent of tebuconazole wettable powderprocessed by Nanjing Nannong Pesticide Technology Development Co., Ltd.on commission which are purchased from the market are adopted. Dosagesof the needed active jinggangmycin ingredient and active tebuconazoleingredient for use are designed according to an experiment, the twotypes of agents are weighed respectively, one type of agent is firstmixed with half of water for dilution, then mixed with the other type ofagent, stirred and then added with the other half of the water, andafter uniform mixing, the mixed agent is sprayed. The mixed agent issprayed for the first time at the initial stage of wheat blooming, andis sprayed for the second time 5 days later, and the amount of sprayedwater per mu is 50 kg. An experiment was carried out in Jiangsu XinyangFarm in April, 2014, the wheat variety was Huaimai 33, each treatmentwas repeated three times, and the area of each plot was 50 squaremeters. A polyketone suspension concentrate was adopted as a controlagent for treatment. According to a corresponding method specified byGuidelines for the Field Efficacy Trials of Fungicides, the occurrencesof wheat scab, powdery mildew, rust and leaf blight were investigated atthe stage of milky ripeness, and disease indexes and control effect werecalculated. Synergistic effects and toxin assay are the same as theabove-mentioned research, and treating dosages and prevention effectscan be seen in Table 10.

TABLE 10 Effect of Jinggangmycin and Tebuconazole Wettable Powder TankMixture in Controlling Wheat Diseases Theoretical Grain Disease IndexPrevention Prevention Toxin Treating Dosage Effect % Effect ContentSerial (g ai. agent/mu) Head Leaf Powdery on Head Synergistic (μg/g ofNumber Jinggangmycin Tebuconazole Blight Blight Mildew Rust Blight %Coefficient kernels)  1 3 15 91.24 92.36 89.26 96.54 56.39 161.80 0.29 2 5 10 90.86 88.41 79.34 98.64 57.33 158.50 0.31  3 5 5 85.41 77.676.49 88.3 51.65 165.36 0.53  4 10 5 89.21 85.64 82.16 89.26 54.91162.47 0.29  5 50 3 83.64 76.34 76.39 76.51 78.75 106.21 0.46  6 100 389.51 86.58 73.11 88.34 52.36 170.96 0.52  7 100 / 31.4 8.5 7.3 5.1 / /2.89  8 50 / 26.7 7.9 4.4 3.6 / / 3.24  9 10 / 12.8 0 4.5 0 / / 3.86 105 / 6.5 0 0 0 / / 4.2 11 3 / 0 0 0 0 / / 4.77 12 / 15 56.39 77.35 72.6188.46 / / 4.02 13 / 10 54.36 72.3 76.19 76.54 / / 3.29 14 / 5 48.2956.49 66.32 70.92 / / 2.14 15 / 3 30.55 35.61 39.46 38.41 / / 2.08 16Polyketone 57 g 69.4 72.1 75.3 74.6 / / 3.16 a.i. 17 Blank Control 45.618.4 20.6 24.8 / / 5.84 Disease Index

The above-mentioned research results indicate that when the singlejinggangmycin agent and the single tebuconazole agent in are mixed in atank on spot and then sprayed, the effect of the treatment using 3 g to100 g of active jinggangmycin ingredient per mu and the effect of thetreatment using 3 g to 15 g of active tebuconazole ingredient per mu incontrolling head blight are equivalent to the effect of a prepared mixedagent applied on barley and wheat, moreover, the synergistic effect onwheat scab, powdery mildew, rust and leaf blight is also obvious, andthe control effect is better than that of polyketone. In particular, thesynergistic effect on wheat scab is prominent, the results are similarto the results of the other examples, which is reflected in that withinthe experimental dosage rang, as the dosage of tebuconazole is reduced,the synergistic effect is enhanced. 3 g to 50 g of active jinggangmyciningredient used per mu and 3 g to 15 g of active tebuconazole ingredientused per mu can both achieve an ideal effect in controlling head blight,controlling leaf blight, powdery mildew and rust and reducing DON toxincontamination.

In a word, Fusarium head blight described in the present invention iswheat scab and barley scab, including bud rot, seedling blight, ear rotor ear blight of grain crops caused by Fusarium pathogenic fungi(Fusarium spp.) of carbendazim-resistant Fusaria.

The above-mentioned research results indicate that the compositioncontaining 1 g to 150 g of active jinggangmycin ingredient and 1 g to 20g of active tebuconazole ingredient used per mu has a remarkablesynergistic effect in treating (coating or mixing) wheat seeds tocontrol Fusarium head blight before sowing, and moreover, thecomposition can also control powdery mildew, rust and sheath blight ofgrain; and when mixed with water and sprayed at the stages of wheatheading, blooming and filling, the composition not only has asynergistic effect for the control of barley scab and wheat scab, butalso has the effect of decreasing the DON toxin contamination level,increasing thousand kernel weight and controlling powdery mildew, rustand leaf blight.

[Beneficial Effect]

The beneficial effect of the present invention is that compared withconventional fungicides, the fungicide composition of the presentinvention has the following advantages: (1) compared with theapplication of the single agents, the composition has an obvioussynergistic effect when used for controlling Fusarium head blight, andcan remarkably enhance the effect of controlling head blight, increasingthe ability of the human being in controlling head blight of graincrops; (2) the contamination level of DON toxin in grain can beremarkably decreased; (3) dosage, cost, environmental pollution andpesticide residue can be reduced; (4) fungicide selectivity isdecreased, the resistance of pathogenic fungi of wheat to tebuconazolecan be delayed, and Fusarium head blight which generates resistance tocarbendazim can be controlled; (5) the persistent period ofjinggangmycin is prolonged, the frequency of application is decreased,the powdery mildew, rust, leaf blight and sheath blight of grain canalso be controlled, and the labor cost of pesticide application isreduced; (6) jinggangmycin is a pollution-free microbial pesticide withhigh selectivity, and by being mixed with tebuconazole, greatly reducesthe dosage of the chemical pesticide, and the composition isenvironment-friendly.

DETAILED DESCRIPTION OF THE INVENTION

The present invention will be better understood through the followingembodiments.

Embodiment 1: Experiment on 40 Percent of Jinggangmycin and TebuconazoleComposition Suspension Concentrate for Controlling Wheat Diseases

Time and Place of Experiment: April, 2015, Jiangsu Xinyang Farm

Experimental Material: wheat, variety: Huaimai 33

Experimental Method:

Sample Preparation Method: Raw tebuconazole and raw jinggangmycin weremixed according to weight ratios of 1:19, 1:7, 1:3, 4:1 and 7:3, andaccording to the method of the research V described in the specificationof the present application, 40 percent of experimental compositionsuspension concentrate samples with different ratios were obtained byprocessing. 25 mL of composition preparation, 50 mL of compositionpreparation and 100 mL of composition preparation were used per mu. Eachcomposition preparation was mixed with water and sprayed at the initialstage of wheat blooming, and was sprayed for the second time 5 dayslater, and the amount of sprayed water per mu was 50 kg. Meanwhile, acorresponding dose of single wettable powder and spraying clean waterwere used as controls, carbendazim served as a control agent, eachtreatment was repeated two times, and the area of each plot was 100square meters.

According to a corresponding method specified by Guidelines for theField Efficacy Trials of Fungicides, the occurrences of wheat scab,powdery mildew, rust and leaf blight were investigated at the stage ofmilky ripeness, disease indexes, control effect, synergistic effect onhead blight and effects in controlling toxin contamination werecalculated, and results are listed in Table 11.

TABLE 11 Effect of 40 Percent of Jinggangmycin and TebuconazoleComposition Suspension Concentrate in Controlling Wheat DiseasesTheoretical Grain Rate of Prevention Toxin Decrease Effect of Content inToxin Mixed (mg of Content Treating Dosage Disease Index PreventionEffect % Agent on DON in Serial (g ai. agent/mu) Head Leaf Powdery HeadSynergistic /Kg of Grains Number Jinggangmycin Tebuconazole BlightBlight Mildew Rust Blight % Coefficient grain) (%)  1 1.0 19.0 91.2 86.598.0 95.5 74.2 122.9 0.32 95.0  2 0.5 9.5 80.2 75.2 81.6 84.5 65.4 122.60.36 94.4  3 2.5 17.5 88.6 83.6 88.5 92.6 73.9 119.9 0.41 93.7  4 1.258.75 76.4 80.5 81.2 86.5 61.5 124.2 0.42 93.5  5 5.0 15.0 86.8 81.3 86.091.3 71.8 120.9 0.45 93.0  6 2.5 7.5 70.4 65.0 65.6 66.7 56.7 124.2 0.3594.6  7 16.0 4.0 71.6 51.6 59.2 54.5 46.3 154.6 0.38 94.1  8 8.0 2.050.2 16.0 31.2 32.1 28.5 176.1 0.68 89.5  9 28.0 12.0 94.2 86.4 88.686.9 77.8 121.1 0.71 89.0 10 14.0 6.0 72.3 70.8 66.9 64.5 56.1 128.80.69 89.3 11 28.0 / 29.6 12.5 12.3 11.2 / / 2.85 55.9 12 16.0 / 27.8 7.911.7 7.6 / / 2.98 53.9 13 14.0 / 23.7 3.2 10.5 0.0 / / 3.45 46.6 14 8.0/ 17.9 2.5 0.0 0.0 / / 3.61 44.1 15 5.0 / 0.0 0.0 0.0 0.0 / / 3.85 40.416 2.5 / 0.0 0.0 0.0 0.0 / / 4.05 37.3 17 1.3 / 0.0 0.0 0.0 0.0 / / 4.1535.8 18 1.0 / 0.0 0.0 0.0 0.0 / / 4.52 30.0 19 0.5 / 0.0 0.0 0.0 0.0 / /5.20 19.5 20 / 19.0 74.2 81.2 81.2 92.8 / / 0.84 87.0 21 / 17.5 73.980.3 76.4 89.4 / / 1.20 81.4 22 / 15.0 71.8 78.9 74.5 85.6 / / 1.64 74.623 / 12.0 68.4 76.5 76.5 81.8 / / 1.84 71.5 24 / 9.5 65.4 74.2 65.2 80.5/ / 2.10 67.5 25 / 8.8 61.5 73.1 60.5 78.4 / / 2.50 61.3 26 / 7.5 56.771.2 58.2 71.5 / / 2.84 56.0 27 / 6.0 42.5 29.5 36.4 45.2 / / 3.24 49.828 / 4.0 25.6 19.2 28.5 32.1 / / 5.12 20.7 29 / 2.0 12.9 14.5 15.4 18.0/ / 5.64 12.7 30 Carbendazim 49.04 / / / / / 4.35 32.7 50 g a.i. 31Blank Control 35.2 14.3 22.7 20.1 / / 6.46 / Disease Index

The above-mentioned field application results indicate that 0.5 g to 28g of jinggangmycin and 2 g to 19 g of tebuconazole have a good wheatdisease control effect when used. The synergistic coefficient for thecontrol of head blight is 120 to 176, so the synergistic effect isprominent. However, when 2.5 g to 28 g of jinggangmycin and 6 g to 19 gof tebuconazole are applied per mu, the toxin contamination level isremarkably decreased, the synergistic effect is extremely obvious, theeffect in controlling head blight is not less than 70 percent, andfurthermore, powdery mildew, rust and leaf blight of wheat can also beeffectively controlled. These results also indicate that when 0.5 g to28 g of jinggangmycin and 2.0 g to 19 g of tebuconazole are adopted permu, the DON toxin content in grains is reduced by not less than 90percent in comparison with that in the control, and in comparison withthe toxin reduction effect of jinggangmycin or tebuconazole used fortreatment alone, the toxin reduction effect is also extremelyoutstanding.

Embodiment 2: Effect of 60 Percent of Jinggangmycin and TebuconazoleComposition Wettable Powder in Controlling Wheat Diseases

Time and Place of Experiment: April, 2015, Jiangsu Yanjiang Institute ofAgricultural Sciences (Rugao)

Experimental Material: wheat. Variety: Yangmai 4

Experimental Method: Raw tebuconazole and raw jinggangmycin were mixedaccording to a weight ratio of 1:1, and 60 percent of experimentalwettable powder sample was prepared according to the method of theresearch VII described in the specification of the present application.The composition wettable powder containing 4 g to 20 g of the activejinggangmycin ingredient and the active tebuconazole ingredient wasrespectively used per mu. The composition wettable powder was mixed withwater and sprayed on Yangmai 4 at the initial stage of wheat blooming,and was sprayed for the second time 5 days later, and the amount ofsprayed water per mu was 45 kg. A dose of 20 percent of single-agentwettable powder used in mixture and spraying clean water were used ascontrols, and 60 g of 50 percent of polyketone wettable powder per muwas adopted as a control agent. Each treatment was repeated three times,and the area of each plot was 50 square meters. According to acorresponding method specified by Guidelines for the Field EfficacyTrials of Fungicides, the occurrences of wheat scab, powdery mildew,rust and leaf blight were investigated at the stage of milky ripeness,and disease indexes and control effect were calculated. The result canbe seen in Table 12.

TABLE 12 Effect of 60 Percent of Jinggangmycin and TebuconazoleComposition Wettable Powder in Controlling Wheat Disease TheoreticalActive Ingredient Disease Index Prevention Prevention Thousand DosageEffect % Effect on Kernel Serial (g ai. agent/mu) Head Leaf Powdery onHead Synergistic Weight Number Jinggangmycin Tebuconazole Blight BlightMildew Rust Blight % Coefficient (g)  1 6 6 86.5 70.6 76.5 76.8 53.4162.0 43.1  2 9 9 90.4 75.2 89.7 83.4 71.8 125.9 42.1  3 15 15 96.1 85.490.5 96.7 78.8 122.0 44.3  4 21 21 96.4 91.5 95.4 98.4 85.0 113.4 44.7 5 21 / 35.4 10.2 8.5 8.9 / / 38.5  6 15 / 28.3 9.5 0 0 / / 40.1  7 9 /17.6 9.4 0 0 / / 37.8  8 6 / 0 0 0 0 / / 41.5  9 / 21 76.8 73.5 81.686.1 / / 42.7 10 / 15 70.4 72.4 76.4 78.3 / / 42 11 / 9 65.8 70.1 70.872.6 / / 41.5 12 / 6 53.4 64.5 62.4 61.5 / / 40.5 13 Polyketone 30 54.665.2 75.4 76.8 / / 39.3 14 Blank Control Disease 35.6 14.8 25.1 20.6 / /37.2 Index

The above-mentioned field application results indicate that thecompositions in which the effective contents of jinggangmycin andtebuconazole according to the weight ratio of 1:1 are 6 g to 21 g eachhas a good control effect on wheat scab, leaf blight, powdery mildew andrust when used per mu, but the effect is optimal when 9 g to 15 g ofactive jinggangmycin ingredient and 9 g to 15 g of active tebuconazoleingredient are used. The composition for treatment can notably increasethousand kernel weight, and the yield-increasing effect is obvious.

Embodiment 3: Effect of 20 Percent of Jinggangmycin and TebuconazoleComposition

Wettable Powder in Controlling Wheat Diseases and Reducing ToxinContamination Time and Place of Experiment: April, 2015, JiangsuYanjiang Institute of Agricultural Sciences (Rugao), arranging theexperiment in the same plots in embodiment 2.

Experimental Material: wheat. Variety: Yangmai 4

Experimental Method: After raw tebuconazole and raw jinggangmycin weremixed according to a weight ratio of 1:1, the mixture was added with 10percent by weight of kaolin carrier, 2 percent by weight of sodiumlignin sulfonate adjuvant, 2 percent by weight ofcarboxymethylcellulose, 1 percent by weight of alkylphenolpolyoxyethylene ether phosphate and 2 percent by weight of polyvinylalcohol and, finally, added with light calcium carbonate carries to 100percent, and the mixture was milled by a sand mill and screened by a300-mesh screen. An experimental composition wettable powder sample, thecontent of which was 20 percent, was prepared. The composition wettablepowders containing 20 g, 15 g and 6 g of the active jinggangmyciningredient and the active tebuconazole ingredient were respectively usedper mu. The composition wettable powder was mixed with water and sprayedon Yangmai 4 at the initial stage of wheat blooming, and was sprayed forthe second time 5 days later, and the amount of sprayed water per mu was45 kg. A dose of 20 percent of single-agent wettable powder used inmixture and spraying clean water were used as controls, and 60 g of 50percent of polyketone wettable powder per mu was adopted as a controlagent. Each treatment was repeated three times, and the area of eachplot was 50 square meters. According to a corresponding method specifiedby Guidelines for the Field Efficacy Trials of Fungicides, theoccurrences of wheat scab, powdery mildew, rust and leaf blight wereinvestigated at the stage of milky ripeness, and disease indexes andcontrol effect were calculated. Experimental results are listed in Table13.

TABLE 13 Effect of 20 Percent of Jinggangmycin and TebuconazoleComposition Wettable Powder in Controlling Wheat Diseases TheoreticalActive Ingredient Disease Index Prevention Effect Prevention PreparationDosage % on Effect Thousand Serial Dosage (g ai. agent/mu) Head LeafPowdery on Head Synergistic Kernel Number (g/mu) JinggangmycinTebuconazole Blight Blight Mildew Rust Blight % Coefficient Weight (g) 1 30 3 3 59.4 52.6 62.6 66.4 32.7 181.7 40.2  2 75 7.5 7.5 88.3 81.586.4 86.7 73.1 120.81 43  3 100 10 10 92.4 83.4 91.4 95.8 81.3 113.5944.6  5 50 10 / 32.4 8.6 12.6 6.8 / / 37.5  6 37.5 7.5 / 24.2 8.8 11.96.5 / / 38.6  7 15 3 / 0 5.8 0 0 / / 37.6  8 50 / 10 72.4 64.5 82.4 82.4/ / 40.8  9 37.5 / 7.5 64.5 68.5 73.5 75.6 / / 41.5 10 15 / 3 32.7 65.468.1 64.5 / / 40 11 60 Polyketone 30 54.6 65.2 75.4 76.8 / / 39.3 12Blank Control Disease Index 35.6 14.8 25.1 20.6 / / 37.2

The above-mentioned experimental results indicate that the jinggangmycinand tebuconazole composition has an outstanding synergistic effect forthe control of head blight and an outstanding yield-increasing effect.Such effects are related to a used dosage, but have no significantrelationship with the content and adjuvant of the processed preparation.

Embodiment 4: Effect of 30 Percent of Jinggangmycin and TebuconazoleComposition Water-dispersible Granule in Controlling Wheat Diseases andReducing Toxin Contamination

Time and Place of Experiment: April, 2015, Jiangsu White Horse Lake Farm

Experimental Material: wheat. Variety: Huamai 4

Experimental Method: 30 percent of composition water-dispersible granulewas obtained by processing according to a weight ratio of 5:1 ofjinggangmycin to tebuconazole. A processing method was as follows: 39.1parts by weight of 64 percent of jinggangmycin (25 parts of activeingredient), 5.3 parts by weight of 95 percent of raw tebuconazole (5parts of active ingredient), 30 parts by weight of ammonium sulfate,17.2 parts by weight of light calcium carbonate, 2.5 parts by weight ofalkylphenol ethoxylates, 2.5 parts by weight of sodium dodecyl benzenesulfonate and 3.5 parts by weight of polyoxyethylene fatty acid wereground, screened and pelletized.

200 g, 100 g, 50 g and 30 g of composition preparations were used permu, mixed with water and sprayed at the initial stage of wheat bloomingand sprayed for the second time 5 days later, and the amount of sprayedwater per mu was 50 kg. 100 g of 20 percent of single wettable powderper mu and spraying clean water were used as controls, each treatmentwas repeated three times, and the area of each plot was 50 squaremeters. According to a corresponding method specified by Guidelines forthe Field Efficacy Trials of Fungicides, the occurrences of wheat scab,powdery mildew, rust and leaf blight were investigated at the stage ofmilky ripeness, and disease indexes and control effect were calculated.Experimental results are listed in Table 14.

TABLE 14 Effect of 30 Percent of Jinggangmycin and TebuconazoleComposition Water- dispersible Granule in Controlling Wheat DiseasesActive Ingredient Dosage Disease Index Prevention Effect % SerialPreparation (g ai. agent/mu) Head Leaf Powdery Number Dosage (g/mu)Jinggangmycin Tebuconazole Blight Blight Mildew Rust 1 200 50 10 91.585.6 94.8 96.8 2 100 25 5 86.5 85.8 91.1 91.5 3 50 12.5 2.5 77.8 86.888.7 81.6 4 30 7.5 1.5 40.6 67.8 75.9 73.6 5 100 20 0 40.2 7.8 10.6 6.56 100 0 20 80.1 84.2 96.2 98.5 7 Blank Control Disease Index 25.9 23.429.7 24.0

The above-mentioned field application effect indicates that when sprayedaccording to 15 g to 60 g of total active ingredients of the compositionwater-dispersible granule with 5 parts of jinggangmycin and 1 part oftebuconazole, the composition water-dispersible granule has a goodcontrol effect on wheat scab, powdery mildew, rust and leaf blight, andin particular, when 15 g to 30 g of total active ingredients of thecomposition is sprayed per mu, the cost performance is best.

Embodiment 5: Effect of 32 Percent of Jinggangmycin and TebuconazoleComposition Emulsion in Water in Controlling Wheat Diseases

Time and Place of Experiment: April, 2015, Jiangsu White Horse Lake Farm

Experimental Material: wheat. Variety: Huamai 4

Experimental Method: A preparation method for 32 percent of compositionemulsion in water was as follows: raw tebuconazole was prepared into asuspension concentrate, and the suspension concentrate was added with anethyleneoxide-propyleneoxide block polymer non-ionic emulsifier andstirred into the uniform oil phase; and in addition, a jinggangmycinaqueous solution, an ethylene glycol antifreezing agent and 1 percent ofbenzoic acid were mixed into the aqueous phase. Under high-speedstirring, the aqueous phase was gradually added into the oil phase, sothat 32 percent of oil-in-water emulsion in water in which the ratio ofjinggangmycin to tebuconazole was 1:3 was prepared.

100 mL, 50 mL, 25 mL and 12.5 mL of composition emulsions in water wereused per mu, each composition emulsion in water was mixed with water andsprayed at the initial stage of wheat blooming and sprayed for thesecond time 5 days later, and the amount of sprayed water per mu was 50kg. 100 g of 20 percent of single wettable powder per mu and sprayingclean water were used as controls, each treatment was repeated threetimes, and the area of each plot was 50 square meters. According to acorresponding method specified by Guidelines for the Field EfficacyTrials of Fungicides, the occurrences of wheat scab, powdery mildew,rust and leaf blight were investigated at the stage of milky ripeness,and disease indexes and control effect were calculated. Experimentalresults are listed in Table 15.

TABLE 15 Effect of 32 Percent of Jinggangmycin and TebuconazoleComposition Emulsion in water in Controlling Wheat Diseases ActiveIngredient Preparation Dosage Disease Index Prevention Effect % SerialDosage (g ai. agent/mu) Head Leaf Powdery Number (mL/mu) JinggangmycinTebuconazole Blight Blight Mildew Rust 1 100 8 24 95.6 84.8 90.8 95.2 250 4 12 90.3 80.1 89.1 93.5 3 25 2 6 74.2 64.1 71.8 72.2 4 12.5 1 3 42.234 42.3 44.2 5 100 20 0 34.2 7.8 10.6 6.5 6 100 0 20 84.6 83.6 88.2 82.47 Blank Control Disease Index 35.9 23.4 29.7 24.6

The above-mentioned field application results indicate that when 8 g to32 g of total active ingredients of the jinggangmycin and tebuconazolecomposition is used per mu, the control effect on wheat scab, powderymildew, rust and leaf blight is good, and when 4 g of active ingredientsof the composition is used per mu, the effect in controlling these wheatdiseases is also outstanding.

Embodiment 6: Effect of 45 Percent of Jinggangmycin and TebuconazoleWettable Powder in Controlling Wheat Diseases and Reducing ToxinContamination

Time and Place of Experiment: April, 2015, Jiangsu Yanjiang Institute ofAgricultural Sciences (Rugao)

Experimental Material: wheat. Variety: Yangmai 4

Experimental Method: 45 percent of experimental wettable powder samplein which the weight ratio of jinggangmycin to tebuconazole was 2:1 wasprepared according to the method of the research VII described in thespecification of the present application, and Jiangsu Yanjiang Instituteof Agricultural Sciences was commissioned to carry out a fieldexperiment. 50 g, 40 g, 30 g and 20 g of composition wettable powderpreparations were used per mu respectively. Each composition wettablepowder preparation was mixed with water and sprayed at the initial stageof wheat blooming, and was sprayed for the second time 5 days later, andthe amount of sprayed water per mu was 50 kg. 20 percent of singlejinggangmycin and tebuconazole wettable powder was adopted, doses ofsingle agents in the used mixed agent and 40 g of active carbendazimingredient were adopted as control agents for treatment, spraying cleanwater was adopted as a blank control, each treatment is repeated threetimes, and the area of each plot was 50 square meters. According to acorresponding method specified by Guidelines for the Field EfficacyTrials of Fungicides, the occurrences of wheat scab and powdery mildewwere investigated at the stage of milky ripeness, and disease indexesand control effect were calculated.

During harvest, two hundred ears were sampled in total every fivetreated points, and after being threshed, the samples were sent toNanjing Agricultural University for toxin content assay. The toxin assaymethod is the same as that mentioned above. Experimental results arelisted in Table 16.

TABLE 16 Effect of 45 Percent of Jinggangmycin and TebuconazoleComposition Wettable Powder in Controlling Wheat Diseases Grain ActiveDisease Synergistic Effect Toxin Ingredient Index TheoreticalTheoretical Content Dosage Prevention Prevention Prevention SynergisticSynergistic mg of Toxin Preparation (g ai. Effect % Effect Effect onCoefficient Coefficient DON/k Reduction Serial Dosage agent/mu) HeadPowdery on Head Powdery on Head on Powdery g of Percentage Number (g/mu)Jinggangmycin Tebuconazole Blight Mildew Blight Mildew Blight Mildewgrain %  1 50 15 7.5 92.5 91.5 77.2 74.8 119.8 122.3 0.35 92.26  2 40 126 90.4 88.6 58.6 67.2 154.3 131.8 0.45 90.04  3 30 9 4.5 74.5 74.7 53.161.6 140.3 121.3 0.68 84.96  4 20 6 3 54.2 58.7 37.1 49.6 146.1 118.31.6 64.60  5 75 15 / 34.2 8.6 / / / / 2.86 36.73  6 60 12 / 10.2 6.6 / // / 3.82 15.49  7 45 9 / 9.4 3 / / / / 4.02 11.06  8 30 6 / 7.4 1.8 / // / 4.14 8.41  9 37.5 / 7.5 65.4 72.4 / / / / 1.89 58.19 10 30 / 6 53.964.9 / / / / 2.14 52.65 11 22.5 / 4.5 48.2 60.4 / / / / 2.4 46.90 12 15/ 3 32.1 48.7 / / / / 2.8 38.05 13 80 g of 50% of 64.6 62.8 / / / / 2.9534.73 Carbendazim Wettable Powder 14 Blank Control 21.8 15.2 / / / /4.52 / Disease Index

It can be seen from the experimental results of the above-mentionedfield application embodiments 1-6 that based on the discovery thatjinggangmycin can inhibit the pathogenic factor DON toxin biosynthesisof the head blight fungi, the jinggangmycin and tebuconazole compositionhas an outstanding synergistic effect for the control of wheat scab whenadopted to treat wheat seeds and be sprayed in fields. Moreover, thecontrol effect on powdery mildew, rust, leaf blight and sheath blight ofgrain crops is also ideal. In addition, after the inventor adopted someof the combined jinggangmycin and tebuconazole compositions to carry outexperimental demonstration in Jiangsu Yanjiang Institute of AgriculturalSciences, Jiangsu Xinyang Farm and White Horse Lake Farm for threeyears, the jinggangmycin and tebuconazole compositions all showed anexcellent prevention effect on barley scab and wheat scab, and couldalso effectively control powdery mildew, leaf blight and rust. Thepresent invention not only solves the major problem confronting China atpresent that the wheat scab fungi are hard to control due to carbendazimresistance, but also can overcome the food safety problem caused bywheat contaminated by DON toxin. Compared with conventional fungicidesin the prior art, the jinggangmycin and tebuconazole composition of thepresent invention can greatly reduce the contamination of grains by theDON toxin of Fusaria, and can also greatly reduce chemical pesticidedosage and application cost because of the synergistic effect, andthereby the jinggangmycin and tebuconazole composition has a goodapplication prospect in reducing environmental pollution, controllingpesticide-resistant fungal diseases of grain and guaranteeing foodsafety.

1-10. (canceled)
 11. A process for administrating a pesticidecomposition on crops, the pesticide composition containing jinggangmycinand tebuconazole in reducing DON toxin biosynthesis, characterized inthat the pesticide composition comprises jinggangmycin and tebuconazoleaccording to the weight ratio of (1-150):(1-30); the DON toxin is causedby fusarium head blight; the fusarium head blight is wheat scab andbarley scab, and the fusarium head blight includes bud rot, seedlingblight, ear rot or ear blight of triticeae crops caused by fusariumpathogenic fungi (Fusarium spp.) of carbendazim-resistant fusarium. 12.The process according to claim 11, characterized in that the pesticidecomposition comprises jinggangmycin and tebuconazole according to theweight ratio of (5-85):(3-24).
 13. The process according to claim 11,characterized in that the pesticide composition comprises jinggangmycinand tebuconazole according to the weight ratio of (5-30):(6-18).
 14. Aprocess for administrating an agricultural fungicide on crops, theagricultural fungicide containing jinggangmycin and tebuconazole inreducing DON toxin biosynthesis, characterized in that the agriculturalfungicide contains 0.1 to 90.0 percent by weight of pesticidecomposition according to any one of claims 11-13, and the balance is alight calcium carbonate carrier and/or an adjuvant chosen from ethyleneglycol, propylene glycol, NNO-1, NNO-7, polyethylene glycol, glycerin,ammonium sulfate, alkylphenol ethoxylates, polyoxyethylene alkyphenolphosphorate or benzoic acid acceptable in pesticides.
 15. The processaccording to claim 14, characterized in that the agricultural fungicidecontains 10 to 75.0 percent by weight of pesticide composition accordingto any one of claims 1-3, and the balance is the carrier and/or theadjuvant acceptable in pesticides.
 16. The process according to claim14, characterized in that the agricultural fungicide contains 26 to 58.0percent by weight of pesticide composition according to any one ofclaims 1-3, and the balance is the carrier and/or the adjuvantacceptable in pesticides.
 17. The process according to claim 14,characterized in that the agricultural fungicide is a suspending agent,an emulsion in water, a microemulsion, a wettable powder or awater-dispersible granule.
 18. A tebuconazole-containing pesticidecomposition for preventing and controlling fusarium head blight and anapplication thereof.